Summary:The microcrystalline cellulose (MCC) was extracted by microwave assisted from cotton wool. The effects of sulfuric acid concentration, time and temperature on the yield were investigated by a single factor experiments and the preparation condition were optimized using 2 3 factorial design and analyzed by statistical test. The obtained MCC were characterized using particle size analyzer, FT-IR and XRD. The results showed that the main parameter in order of increasing effects on the MCC yield were time, temperature and acid concentration. The optimum condition was found at acid concentration 55% (w/w), time 1 minute and temperature 30C. The range of particle size at that condition was from 0.052 mm to 353.078 mm, the yield was 78.67% (w/w) and the crystallinity index was 53.16%. Moreover the crystal structure transformed from cellulose I to cellulose II. The crystal structure of cellulose II was obtained without mercerization and ionic liquid treatments.